2 780 Zick, Meyer, von Schütz, Holle, Dwenger, Mitzkat and Hürter: Insulin binding to erythrocytes in children with type-i diabetes K a ± s Ka (10 8 1/mol) X 0 ± s Xo (nmol/1) a 11,6 ±7,6 0,068 ± 0,028 b 19,3 ± 10,0 0,043 ± 0,030 24,0 ± 4,5 0,029 ± 0,007 Die Resultate zeigen, daß die Umstellung von Sehweineihsulin auf biosynthetisches Humaninsulin sowohl kurz- als auch langfristig eine Zunahme der Affinität und eine Abnahme der Konzentration der erythrocytären Insulinrezeptoren bewirkt. Introduction With the introduction of biosynthetic human insulin into diabetes therapy, the question of the biological activity of this new insulin arises, in particular in comparison with the insulin pieparations used so far. When analysing the insulin effect, the first step consists in the binding to specific receptors on the cell surface of the single target organs (1,2). This coupling may be modulated by short-term change in the receptor affinity and by long-term Variation of the receptor conceniration (3,4). In the in vitro studies to date, which only included short-term alterations, no differences between the binding properties of biosynthetic human insulin and other insulins were revealed (5, 6, 7). Therefore it was the objective of our investigations to detect in vivo predominantly longer-term changes in the binding behaviour of insulin to receptors after switching from pork insulin to biosynthetic human insulin. The model of the erythrocyte insulin receptor was selected, because the erythrocyte is subject to receptor alterations similar to those occurring in the peripheral tissues, and the small blood völume reqüired can be technically and psychologically justified in clinical studies. The following data are the results of investigations performed one and five months after switching the insulin preparation. Prior to the change (group a) all children had received an. iridividual raixture of regulär and NPH porcine insulin (Nordisk). They were transferred to an individual mixture of regulär and NPH biosynthetic human insulin (Lilly); The binding studies were repeated one month (group b).as well äs 5 months (group c) after ehanging the insulin preparation. The body height of all children was between the 25th and 75th percentile; all children had normal weight corresponding to their heights. Liver diseases and renal diseases äs well äs disturbances of blood formation had been excluded by preliminary examinatioris. Insulin binding studies 12 hours after the last insulin injectioh and the last food ingestion, approximately 10 ml of heparinized whole blood were taken by venipuncture. Insulin binding was performed according to the procedure of Gambhir (9) with slight modifications (10). Isolated erythrocytes ( x 10 I2 /1) were incubated with porcine [^25 i]insulin (0.05 nmol/1) and unlabeled porcine insulin ( nmol/1) at 15 C for 3 hours. Analytical procedures Free insulin and daily C-peptide excretion in urine wefe determined radioimmunologically (11, 12). HbA,i c values in blood were measured according to I.e. (13). Calculation procedures For parariieter extraction a nonlinear regression procedure was applied which is based on an algorithm indicated by Marquardt (14). The insulin receptor affinity (K a ), the insulin receptor concentration (Xo), and the nonspecific binding (U) were iteratively determined for the one-class binding model via the minimization of the sum of the squares of erirpr with the aid of a HP A Computer. For statistical analyses, parametric procedures (t-test for paired data) were applied in cases of normal distributioh of the data; otherwise nonparametric procedures (Wilcoxon^iest) were employed. ' : ' Materials and Methods Subjects Insulin binding to erythrocytes was measured in 27 children ($ 18, 6 9) aged between 12 and 18 years ( ± s, ± 1.76 years) with type-i-diabetes. The duration of diabetes was between 2 7 und 14.9 years (x ± s> 7.48 ± 2.19 years), and the mean age of diabetes manifestation was 8.2 years. All children received two insulin injections per day, and the daily insulin requirement was more than 0.5 lu/kg body weight (0.86 ± 0.23, x ± s); i.e., all patients were in a phase of postremission defmed by Akerbloom (8). Results One month after changing from porcine insulin to biosynthetic human insulin the insulin receptor affinity had significantty increased from 11.6 to 19.3 x /mol (p < 0,001). After another period of 4 months the receptor affinity increased to 24.0 x l O 8 1/mol, thus demonstrating an incfease of 105.7% in comparison with the receptor affinity priof to switching the insulin therapy (fig. l, tabs. l and 2). >? J. Clin. Chem. Clin. Biochem. / Vol. 21,1983 / No. 12

4 W DE G Walter de Gruyter Berlin-New York J. Büttner (Editor) History of Clinical Chemistry crn 26 cm. 91 pages with illustrations. Hardcover. DM 98,-; approx. US $44.75 ISBN 311 OÖ8Ö122 Clinical Chemistry is a special dlsciplihe of fnedicine which, due to its close relationship both to medicine and to chemistry, is of particular interest to the historian of science. This "History of Clinical Chemistry" is based on a modern outlopk on the history of sciehce. Since the investigatiqn of the histöry of clinical chemistry is still in progress, the book is divided into eight separate contributions, written primarily by historians of science, which togethef prpvide a good coverage of the history öf Clinical Chemistry in the nineteenth Century. The book is written entirely in English and will therefore appeal to an international readership. Each contributioh is prövided with numerous notes and references. Contents Johannes Büttner Introduction Nikolaus Mari/ The historical background of Clinical Chemistry Joseph S. Frütpn - Biochemistry and Clinical Chemistry. A retrospect Erika Hickel - The emergence of Clinical Chemistry in the 19th Century. Presuppositions and consequences Johannes Büttner Johann Joseph von Scherer ( ). A commentary on the early history of Clinical Chemistry Hans H. Simmer Medicihe and Chemistry around the middle of the 19th Century in Erlangen: Eugen Franz Freiherr von Gofup-Besanez ( ) Johannes Büttner Evolution of Clinical Enzymolögy Johannes Büttner - Relationships between Clinical Medicihe ähd Clinical Chemistry, illustrated by the example of the German-speaking countries in the late 19th Century Wendeil T. Caraway Major developments in clinical Chemical Instrumentation. Prtces are subject to change without notice (HO)

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